Search results for: protein metabolism
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2693

Search results for: protein metabolism

1253 Identification and Characterization of Inhibitors of Epoxide Hydrolase from Trichoderma reesei

Authors: Gabriel S. De Oliveira, Patricia P. Adriani, Christophe Moriseau, Bruce D. Hammock, Felipe S. Chambergo

Abstract:

Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have high biotechnological interest for the drug design and chemistry transformation for industries. In this study, we describe the identification of substrates and inhibitors of epoxide hydrolase enzyme from the filamentous fungus Trichoderma reesei (TrEH), and these inhibitors showed the fungal growth inhibitory activity. We have used the cloned enzyme and expressed in E. coli to develop the screening in the library of fluorescent substrates with the objective of finding the best substrate to be used in the identification of good inhibitors for the enzyme TrEH. The substrate (3-phenyloxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester showed the highest specific activity and was chosen for the next steps of the study. The inhibitors screening was performed in the library with more than three thousand molecules and we could identify the 6 best inhibitors. The IC50 of these molecules were determined in nM and all the best inhibitors have urea or amide in their structure, because It has been recognized that these groups fit well in the hydrolase catalytic pocket of the epoxide hydrolases. Then the growth of T. reesei in PDA medium containing these TrEH inhibitors was tested, and fungal growth inhibition activity was demonstrated with more than 60% of inhibition of fungus growth in the assay with the TrEH inhibitor with the lowest IC50. Understanding how this EH enzyme from T. reesei responds to inhibitors may contribute for the study of fungal metabolism and drug design against pathogenic fungi.

Keywords: epoxide hydrolases, fungal growth inhibition, inhibitor, Trichoderma reesei

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1252 Stimulation of Nerve Tissue Differentiation and Development Using Scaffold-Based Cell Culture in Bioreactors

Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

Abstract:

Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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1251 The Effects of Varying Nutrient Conditions on Hydrogen Production in PGR5 Deficient C. Reinhardtii Mutants

Authors: Samuel Mejorado

Abstract:

C. Reinahrdtii serves as one of the most promising organisms from which to obtain biological hydrogen. However, its production catalyst, [FeFe]-hydrogenase, is largely inhibited by the presence of oxygen. In recent years, researchers have identified a Proton Gradient Regulation 5 (PGR5) deficient mutant, which shows enhanced respiration and lower accumulations of oxygen within the system. In this research, we investigated the effects of varying nutrient conditions on PGR5 mutants' ability to produce hydrogen. After growing PGR5 mutants in varying nutrient conditions under 55W fluorescent lamps at 30℃ with constant stirring at 200 rpm, a common water displacement method was utilized to obtain a definitive volumetric reading of hydrogen produced by these mutants over a period of 12 days. After the trials, statistical t-tests and ANOVAs were performed to better determine the effect which nutrient conditions have on PGR5 mutants' ability to produce hydrogen. In this, we report that conditions of sulfur deprivation most optimally enhanced hydrogen production within these mutants, with groups grown under these conditions demonstrating the highest production capacity over the entire 12-day period. Similarly, it was found that when grown under conditions of nitrogen deprivation, a favorable shift towards carbon fixation and overall lipid/starch metabolism was observed. Overall, these results demonstrate that PGR5-deficient mutants stand as a promising source of biohydrogen when grown under conditions of sulfur deprivation. To date, photochemical characteristics of [FeFe]-hydrogenase in these mutants have yet to be investigated under conditions of sulfur deprivation.

Keywords: biofuel, biohydrogen, [FeFe]-hydrogenase, algal biofuel

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1250 Association of Xeroderma pigmentosum Group D Gene Polymorphism with Colorectal Cancer Risk in Kashmiri Population

Authors: Syed Sameer Aga, Saniya Nissar

Abstract:

The Xeroderma pigmentosum group D gene (XPD) plays a key role in nucleotide excision repair (NER) pathway of the damaged DNA. Genetic polymorphisms in the coding region of the XPD gene may alter DNA repair capacity of the protein and hence can modulate the risk of colorectal cancer (CRC) risk. The aim of the study was to determine the genetic association of XPD Lys751Gln polymorphism with the risk of colorectal cancer (CRC) development. 120 CRC patients and 160 normal controls were assessed for genotype frequencies of XPD Lys751Gln polymorphism using PCR-RFLP technique. We observed a significant association (p < 0.05) between the XPD Lys751Gln polymorphism and the risk of developing CRC (p < 0.05). Additionally, Gln/Gln genotype of the XPD gene doubled the risk for the development of CRC [p < 0.05; OR=2.25 95% CI (1.07-4.7)]. Our results suggest that there is a significant association between the XPD Lys751Gln polymorphism and the risk of CRC.

Keywords: colorectal cancer, polymorphism, RFLP, DNA Repair, NER, XPD

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1249 The Decrease of Collagen or Mineral Affect the Fracture in the Turkey Long Bones

Authors: P. Vosynek, T. Návrat, M. Peč, J. Pořízka, P. Diviš

Abstract:

Changes of mechanical properties and response behavior of bones is an important external sign of medical problems like osteoporosis, bone remodeling after fracture or surgery, osteointegration, or bone tissue loss of astronauts in space. Measuring of mechanical behavior of bones in physiological and osteoporotic states, quantified by different degrees of protein (collagen) and mineral loss, is thus an important topic in biomechanical research. This contribution deals with the relation between mechanical properties of the turkey long bone–tibia in physiological, demineralized, and deproteinized state. Three methods for comparison were used: densitometry, three point bending and harmonic response analysis. The results help to find correlations between the methods and estimate their possible application in medical practice.

Keywords: bone properties, long bone, osteoporosis, response behavior

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1248 Construction of a Fusion Gene Carrying E10A and K5 with 2A Peptide-Linked by Using Overlap Extension PCR

Authors: Tiancheng Lan

Abstract:

E10A is a kind of replication-defective adenovirus which carries the human endostatin gene to inhibit the growth of tumors. Kringle 5(K5) has almost the same function as angiostatin to also inhibit the growth of tumors since they are all the byproduct of the proteolytic cleavage of plasminogen. Tumor size increasing can be suppressed because both of the endostatin and K5 can restrain the angiogenesis process. Therefore, in order to improve the treatment effect on tumor, 2A peptide is used to construct a fusion gene carrying both E10A and K5. Using 2A peptide is an ideal strategy when a fusion gene is expressed because it can avoid many problems during the expression of more than one kind of protein. The overlap extension PCR is also used to connect 2A peptide with E10A and K5. The final construction of fusion gene E10A-2A-K5 can provide a possible new method of the anti-angiogenesis treatment with a better expression performance.

Keywords: E10A, Kringle 5, 2A peptide, overlap extension PCR

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1247 Impact of Edible Coatings Made of Chitosan and Spray Dried Propolis in the Shell Life of White Cachama (Piaractus brachypomus)

Authors: David Guillermo Piedrahita Marquez, Hector Suarez Mahecha, Jairo Humberto Lopez

Abstract:

There is a need to preserve aquaculture matrices due to their high nutritional value, and its broad consumption, one of those species is the white cachama (Piaractus brachypomus), this fish is located in the rivers of eastern Colombia, and the previously mentioned species needs more study. Therefore, in a paper the effects of an alternative method of preservation of shell life were investigated, the method used is the application of an edible coating made from chitosan and ethanolic extract of propolis (EEP) encapsulated in maltodextrin. The coating was applied by immersion, and after that, we investigated the post mortem quality changes of the fish performing physicochemical and microbiological analysis. pH, volatile bases, test thiobarbituric acid and peroxide value were tested; finally, we studied the effect of the coating on mesophilic strains, coliforms and other microorganisms such as Staphylococcus, and Salmonella. Finally, we concluded that the coating prolongs the shelf life because it acts as a barrier to oxygen and moisture, the bioactive compounds trap free radicals and the coatings changes the metabolism and cause the cell lysis of the microorganisms. It was determined that the concentration of malonaldehyde, the volatile basic nitrogen content and pH are the variables that distinguish more clearly between the samples with the treatment and the control samples.

Keywords: antimicrobial activity, lipid oxidation, texture profile analysis (TPA), sensorial analysis, peroxide value, thiobarbituric acid assay (TBA), total volatile basic nitrogen (TVB-N)

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1246 Introduction of PMMA-Tag to VHH for Improving Recovery and Immobilization Rate of VHHS

Authors: Bongmun Kang, Kagnari Yamakawa, Yoshihisa Hagihara, Yuji Ito, Michimasa Kishimoto, Yoichi Kumada

Abstract:

The PMMA-tag was genetically fused with the C-terminal region of VHH molecules. This antibody, VHH, is known as a single-chain domain, which is devoid of light chains. The PMMA-tag, which could affect the isoelectric point (pI) changeable with a charge of amino acid in VHHs were closely related to the solubility of VHH molecules during refolding. The genetic fusion of PMMA-tag to C-terminal region of VHHs significantly affects the recovery of their soluble protein during refolding by 50 mM TAPS at pH 8.5. It could be refolded with a recovery of more than 95% by dialysis at pH 8.5. A marked difference in the antigen-binding activities in the adsorption state was significantly high in VHH-PM compared to the wild type of VHH. There are approximately 8-fold differences in the antigen-binding activities in the adsorption state between VHH-PM and VHH.

Keywords: VHH, PMMA-tag, isoelectric point, pH, Solubility, refolding, immobilization, ELISA

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1245 A Green Approach towards the Production of CaCO₃ Scaffolds for Bone Tissue Engineering

Authors: Sudhir Kumar Sharma, Abiy D. Woldetsadik, Mazin Magzoub, Ramesh Jagannathan

Abstract:

It is well known that bioactive ceramics exhibit specific biological affinities, especially in the area of tissue re-generation. In this context, we report the development of an eminently scalable, novel, supercritical CO₂ based process for the fabrication of hierarchically porous 'soft' CaCO₃ scaffolds. Porosity at the macro, micro, and nanoscales was obtained through process optimization of the so-called 'coffee-ring effect'. Exposure of these CaCO₃ scaffolds to monocytic THP-1 cells yielded negligible levels of tumor necrosis factor-alpha (TNF-α) thereby confirming the lack of immunogenicity of the scaffolds. ECM protein treatment of the scaffolds showed enhanced adsorption comparable to standard control such as glass. In vitro studies using osteoblast precursor cell line, MC3T3, also demonstrated the cytocompatibility of hierarchical porous CaCO₃ scaffolds.

Keywords: supercritical CO2, CaCO3 scaffolds, coffee-ring effect, ECM proteins

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1244 SEC-MALLS Study of Hyaluronic Acid and BSA Thermal Degradation in Powder and in Solution

Authors: Vasile Simulescu, Jakub Mondek, Miloslav Pekař

Abstract:

Hyaluronic acid (HA) is an anionic glycosaminoglycan distributed throughout connective, epithelial and neural tissues. The importance of hyaluronic acid increased in the last decades. It has many applications in medicine and cosmetics. Hyaluronic acid has been used in attempts to treat osteoarthritis of the knee via injecting it into the joint. Bovine serum albumin (also known as BSA) is a protein derived from cows, which has many biochemical applications. The aim of our research work was to compare the thermal degradation of hyaluronic acid and BSA in powder and in solution, by determining changes in molar mass and conformation, by using SEC-MALLS (size exclusion chromatography -multi angle laser light scattering). The aim of our research work was to observe the degradation in powder and in solution of different molar mass hyaluronic acid samples, at different temperatures for certain periods. The degradation of the analyzed samples was mainly observed by modifications in molar mass.

Keywords: thermal degradation, hyaluronic acid, BSA, SEC-MALLS

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1243 Differentially Expressed Protein Biomarkers in Early and Advanced Stage Young Triple-Negative Breast Cancer Patients

Authors: Shamim Mushtaq, Moazzam Shahid

Abstract:

Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.

Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA

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1242 Atomic Force Microscopy Studies of DNA Binding Properties of the Archaeal Mini Chromosome Maintenance Complex

Authors: Amna Abdalla Mohammed Khalid, Pietro Parisse, Silvia Onesti, Loredana Casalis

Abstract:

Basic cellular processes as DNA replication are crucial to cell life. Understanding at the molecular level the mechanisms that govern DNA replication in proliferating cells is fundamental to understand disease connected to genomic instabilities, as a genetic disease and cancer. A key step for DNA replication to take place, is unwinding the DNA double helix and this carried out by proteins called helicases. The archaeal MCM (minichromosome maintenance) complex from Methanothermobacter thermautotrophicus have being studied using Atomic Force Microscopy (AFM), imaging in air and liquid (Physiological environment). The accurate analysis of AFM topographic images allowed to understand the static conformations as well the interaction dynamic of MCM and DNA double helix in the present of ATP.

Keywords: DNA, protein-DNA interaction, MCM (mini chromosome manteinance) complex, atomic force microscopy (AFM)

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1241 Metabolic Costs and Chemical Profiles of Wax Production in Cryptolaemus montrouzieri and Tenuisvalvae notata

Authors: Nataly De La Pava, Christian S. A. Silva-Torres, Arodí P. Favaris, José Maurício S. Bento

Abstract:

The lady beetles Tenuisvalve notata and Cryptolaemus montrouzieri are important predators of mealybugs (Hemiptera: Pseudococcidae). Similar to the prey, these lady beetles produce wax filaments that cover their body during the larval stage. It has been hypothesized that lady beetle body wax chemical profiles are similar to their prey as i) a mechanism of camouflage and ii) conveying protection to the lady beetle larvae against aphid-tending predatory ants. In this study, we tested those hypotheses for the predators T. notata and C. montrouzieri and two mealybug prey species, Ferissia dasyrilii, and Planococcus citri. Next, we evaluated the influence of feeding on cuticular chemistry during predator development and identified possible metabolic costs associated with wax production. Cuticular wax samples were analyzed by GC-MS and GC-FID. Also, the metabolic cost linked to wax production was evaluated in the 4th instar larvae of the two predators when subjected to body wax removal from 0 to 4 times. Results showed that predator body wax profiles are not similar to the chemical profile of prey body wax. There was a metabolic cost associated with wax removal; predators (male and female) showed a significant reduction in adult body weight when the wax was removed. This suggests the reallocation of energy to wax replacement instead of growth. In addition, it was detected effects of wax removal on fecundity and egg viability. The results do not support the hypothesis that predators mimic the cuticular wax composition of prey as a means of camouflage.

Keywords: biological control, body wax, coccinellids, cuticular hydrocarbons, metabolism cost, reproduction

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1240 The Proportion of and Factors Associated With Thyroid Dysfunction among Individuals Referred To A Tertiary Care Facility in Kabul, Afghanistan

Authors: Mohammad Naeem Lakanwall

Abstract:

Background:The thyroid gland, located just below the vocal cord on each side of and anterior to the trachea, is one of the main endocrine glands. Its normal weight is 15 to 20 grams in adults. The thyroid secretes two most important hormones, thyroxine and triiodothyronine, usually called T4 and T3, respectively. These hormones greatly increase the metabolic rate of the body. In addition to T3 and T4, the thyroid gland secrets calcitonin as well which is a significant hormone for calcium metabolism. Objective: The aim of this study is to estimate the proportion of and to identify factors associated with thyroid dysfunction among individuals coming to a tertiary care facility in Kabul, Afghanistan. Material and Methods: An analytical cross-sectional study design was conducted from July to Sep 2018. Blood samples were obtained, serum TSH levels were measured, and the patients were divided into three diagnostic categories according to their serum TSH concentrations. 1) Hypothyroidism 2) Hyperthyroidism 3) Normal thyroid Results: A total of 127 individuals were part of the sample for the final analysis. The majority of study participants (77%) were females. A large number of the participants (92%) did not have a family history of thyroid dysfunction and the majority of the female participants, (85%) were not pregnant in the last two years. Furthermore, 98% of participants, were non-smokers. Conclusion: The findings of the current study showed a high prevalence of thyroid dysfunctions in individuals coming to FMIC for thyroid functions tests. The findings also indicated that aging and smoking are the factors associated with thyroid dysfunctions. Further studies are needed to find out the prevalence of and factors associated with thyroid dysfunctions.

Keywords: Afghanistan, Kabul, hypothyroidism, hyperthyroidism, triiodothyronine, thyroxine

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1239 Chemical Characterization and Prebiotic Effect of Water-Soluble Polysaccharides from Zizyphus lotus Leaves

Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

Abstract:

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% are neutral sugar and the rest 16.25±1.62% are uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructo-oligosaccharide (RP95). The oligosaccharides concentration was 1g/L of man rogosa sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects

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1238 Dys-Regulation of Immune and Inflammatory Response in in vitro Fertilization Implantation Failure Patients under Ovarian Stimulation

Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

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Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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1237 A Review on the Importance of Nursing Approaches in Nutrition of Children with Cancer

Authors: Ş. Çiftcioğlu, E. Efe

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In recent years, cancer has been at the top of diseases that cause death in children. Adequate and balanced nutrition plays an important role in the treatment of cancer. Cancer and cancer treatment is affecting food intake, absorption and metabolism, causing nutritional disorders. Appropriate nutrition is very important for the cancerous child to feel well before, during and after the treatment. There are various difficulties in feeding children with cancer. These are the cancer-related factors. Other factors are environmental and behavioral. As health professionals who spend more time with children in the hospital, nurses should be able to support the children on nutrition and help them to have balanced nutrition. This study aimed to evaluate the importance of nursing approaches in the nutrition of children with cancer. This article is planned as a review article by searching the literature on this field. Anorexia may develop due to psychogenic causes or chemotherapeutic agents or accompanying infections and nutrient uptake may be reduced.  In addition, stomatitis, mucositis, taste and odor changes in the mouth, the feeling of nausea, vomiting and diarrhea can also reduce oral intake and result in significant losses in the energy deficit. In assessing the nutritional status of children with cancer, determining weight loss and good nutrition is essential anamnesis of a child.  Some anthropometric measurements and biochemical tests should be used to evaluate the nutrition of the child. The nutritional status of pediatric cancer patients has been studied for a long time and malnutrition, in particular under nutrition, in this population has long been recognized. Yet, its management remains variable with many malnourished children going unrecognized and consequently untreated. Nutritional support is important to pediatric cancer patients and should be integrated into the overall treatment of these children.

Keywords: cancer treatment, children, complication, nutrition, nursing approaches

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1236 Liver Histopathological Findings after Treatment with Anastrazole and Letrozole in Ovariectomized Rats

Authors: Ioannis Boutas, Vasilios Pergialiotis, Nicolaos Salakos, George Agrogiannis, Panagiotis Konstantopoulos, Laskarina-Maria Korou, Theodoros Kalampokas, Odysseas Gregoriou, George Creatsas, Despina Perrea

Abstract:

Introduction: The effect of third generation aromatase inhibitors in the lipid profile among women with breast cancer, present diversities. It has been also shown that low levels of estrogens affect liver metabolism in mice in numerous ways, such as lipid accumulation and hepatic steatosis. Materials and Methods: Forty-five female Wistar rats underwent surgical ovariectomy. The animals were anesthetized with a combination of ketamine (75 mg/kg) and xylazine (10 mg/kg) which were administered intraperitoneally. After the ovariectomy, the operated animals were randomized in three groups. The first group did not receive any drug regimen (ovariectomized control group). The second group received Anastrazole and the third group received Letrozole. Four months after the initiation of the study, the animals were euthanized and livers were dissected immediately for further histopathological analysis. The histological features were grouped into 4 broad categories: steatosis, ballooning, portal inflammation and lobular activity. A score from 0 (absence) to 3 (severe) was assigned to each parameter. Results: The liver pathology analysis revealed significant differences among groups with favored mild steatosis and ballooning among animals that received Anastrazole or Letrozole. Conclusion: The effect of Anastrazole and Letrozole on liver function have not yet been clarified. In our study mild histological liver alterations seem also to occur and these alterations should be taken in mind in future clinical studies

Keywords: anastrazole, letrozole, liver, rats

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1235 Body Composition Evaluation among High Intensity and Long Term Walking Distance Participants

Authors: Priscila Vitorino, Jeeziane Rezende, Edison Pereira, Adrielly Silva, Weimar Barroso

Abstract:

Body composition insight during physical activity is relevant to follow up sports income since it can be important and actuate in velocity, resistance, potency, and has an effect on force and agility. The purpose of this study was to identify anthropometric profile, evaluate and correlate body mass index and bioimpedance behavior during the days of Caminhada Ecológica de Goiás - Brasil. A longitudinal study was performed with 25 male participants, with an average age of 45.6±9.1 years. All patients were actives. Body composition was evaluated by body mass index (BMI) measurement and bioimpedance procedures. Both were collected 20 days before walking beginning (A0) and in the four days along the same (A1, A2, A3 e A4). Data were collected in the end of each walking day at athletes accommodations. Final distance during walking route was 308 km in five days, with an average of 62km/day and 7,6 km/hour, and an average temperature of 30°C. Data are represented with mean and standard deviation. ANOVA (Bonferroni pos test) was used to compare frequent measurements between the days. Pearson's correlation test was used to correlate BMI with lean mass, fat mass, and water. BMI decreased from A0 to A1, A2 and A3 (p < 0,01) and increased on A4 (p < 0,01). No changes were observed concerning fat percentage (p=0,60), lean mass (p=0,10) and body water composition (p=0,09). A positive and moderate correlation between BMI and fat percentage was observed; an inverse and moderate correlation between BMI, lean mass and body water composition occurred. Total body mass increased during high intensity and long term walking distance. However, the values of body fat, lean mass and water were maintained.

Keywords: aerobic exercise, body composition, metabolism, sports

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1234 Mechanisms of O-1602 Induced Endothelium-Independent Vasorelaxation of Rat Small Mesenteric Artery

Authors: Yousuf Al Suleimani, Ahmed Al Mahruqi

Abstract:

A typical cannabinoid O-1602 induces vasorelaxation and activates the orphan G protein-coupled receptor GPR55 in human endothelial cells. The aim of this study is to characterize the mechanisms of endothelium-independent relaxation of O-1602 in the rat small mesenteric artery using wire myograph. In endothelium-denuded vessels, O-1602 partially produced concentration-dependent vasorelaxation. In vessels depleted of intracellular Ca2+ (by EGTA and methoxamine), CaCl2 produced concentration-dependent contraction. Preincubation with O-1602 (at 10 µM and 30 µM) abolished the contractile responses (P<0.01). The putative antagonist at novel “endothelial anandamide receptor” O-1918 (10 µM) significantly reversed the inhibitory effect of O-1602 on CaCl2-induced vasoconstriction. It is likely that the mechanism of endothelium-independent vasorelaxation to O-1602 is mediated by interfering with Ca2+ entry via an O-1918-sensitive pathway.

Keywords: O-1602, endothelium, vasorelaxation, calcium

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1233 Association of Zinc with New Generation Cardiovascular Risk Markers in Childhood Obesity

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Zinc is a vital element required for growth and development. This fact makes zinc important, particularly for children. It maintains normal cellular structure and functions. This essential element appears to have protective effects against coronary artery disease and cardiomyopathy. Higher serum zinc levels are associated with lower risk of cardiovascular diseases (CVDs). There is a significant association between low serum zinc levels and heart failure. Zinc may be a potential biomarker of cardiovascular health. High sensitive cardiac troponin T (hs-cTnT) and cardiac myosin binding protein C (cMyBP-C) are new generation markers used for prediagnosis, diagnosis, and prognosis of CVDs. The aim of this study is to determine zinc as well as new generation cardiac markers profiles in children with normal body mass index (N-BMI), obese (OB), morbid obese (MO) children, and children with metabolic syndrome (MetS) findings. The association among them will also be investigated. Four study groups were constituted. The study protocol was approved by the institutional Ethics Committee of Tekirdag Namik Kemal University. Parents of the participants filled informed consent forms to participate in the study. Group 1 is composed of 44 children with N-BMI. Group 2 and Group 3 comprised 43 OB and 45 MO children, respectively. Forty-five MO children with MetS findings were included in Group 4. World Health Organization age- and sex-adjusted BMI percentile tables were used to constitute groups. These values were 15-85, 95-99, and above 99 for N-BMI, OB, and MO, respectively. Criteria for MetS findings were determined. Routine biochemical analyses, including zinc, were performed. High sensitive-cTnT and cMyBP-C concentrations were measured by kits based on enzyme-linked immunosorbent assay principle. Appropriate statistical tests within the scope of SPSS were used for the evaluation of the study data. p<0.05 was accepted as statistically significant. Four groups were matched for age and gender. Decreased zinc concentrations were measured in Groups 2, 3, and 4 compared to Group 1. Groups did not differ from one another in terms of hs-cTnT. There were statistically significant differences between cMyBP-C levels of MetS group and N-BMI as well as OB groups. There was an increasing trend going from N-BMI group to MetS group. There were statistically significant negative correlations between zinc and hs-cTnT as well as cMyBP-C concentrations in MetS group. In conclusion, inverse correlations detected between zinc and new generation cardiac markers (hs-TnT and cMyBP-C) have pointed out that decreased levels of this physiologically essential trace element accompany increased levels of hs-cTnT as well as cMyBP-C in children with MetS. This finding emphasizes that both zinc and these new generation cardiac markers may be evaluated as biomarkers of cardiovascular health during severe childhood obesity precipitated with MetS findings and also suggested as the messengers of the future risk in the adulthood periods of children with MetS.

Keywords: cardiac myosin binding protein-C, cardiovascular diseases, children, high sensitive cardiac troponin T, obesity

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1232 Effect of Dust Rejected by Iron and Steel Complex on Roots of Bean Phaseolus vulgaris

Authors: Labiba Zerari Bourafa, Djebar Mohamed Reda, Berrebah Houria, Khadri Sihem, Chiheb Linda

Abstract:

The study of the effect of metal dust (pollutants) was performed on higher plant white beans Phaseolus vulgaris; the experience took place in cellular toxicology laboratory (in vitro culture). The seeds of the bean Phaseolus vulgaris are cultured in a metal contaminated dust medium (a single treatment by different increasing doses), at a rate of 10 seeds per box, for 10 days. The measurement of morpho-metric parameters is performed during the first 96 hours that follow the germination; while the dosage of the proline, the protein content and histological sections are formed on the tenth day (240 h). All morpho-metric and biochemical parameters measured were highly disturbed by metal dust; histological sections confirm this disurbance.

Keywords: conductive fabrics, metal dust, osmoticums, roots, Phaseolus vulgaris

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1231 Activation of TNF-α from Human Endothelial Cells by Exposure of the Mitochondrial Stress Protein (Hsp60) Secreted from THP-1 Monocytes to High Glucose

Authors: Ryan D. Martinus

Abstract:

Inflammation of the endothelium is an important process leading to diabetic atherosclerosis. However, the molecular mechanisms by which diabetes contributes to endothelial inflammation remain to be established. Using In-vitro cultured Human cells and Hsp60 specific ELISA assays, we show that Hsp60 is not only induced in Human monocyte cells under hyperglycaemic conditions but that the Hsp60 is also secreted from these cells. Furthermore, we also demonstrate that the Hsp60 secreted from these monocyte cells is also able to activate Toll-like receptor-4 (TLR4) from Human endothelial cells. This suggests that a potential link may exist between the hyperglycaemia-induced expression of Hsp60 in monocyte cells and vascular inflammation. Circulating levels of Hsp60 due to mitochondrial stress in diabetes patients could, therefore, be an important modulator of inflammation in endothelial cells and thus contribute to the increased incidences of atherosclerosis in diabetes mellitus.

Keywords: mitochondria, Hsp60, inflammation, diabetes mellitus

Procedia PDF Downloads 177
1230 Improved Signal-To-Noise Ratio by the 3D-Functionalization of Fully Zwitterionic Surface Coatings

Authors: Esther Van Andel, Stefanie C. Lange, Maarten M. J. Smulders, Han Zuilhof

Abstract:

False outcomes of diagnostic tests are a major concern in medical health care. To improve the reliability of surface-based diagnostic tests, it is of crucial importance to diminish background signals that arise from the non-specific binding of biomolecules, a process called fouling. The aim is to create surfaces that repel all biomolecules except the molecule of interest. This can be achieved by incorporating antifouling protein repellent coatings in between the sensor surface and it’s recognition elements (e.g. antibodies, sugars, aptamers). Zwitterionic polymer brushes are considered excellent antifouling materials, however, to be able to bind the molecule of interest, the polymer brushes have to be functionalized and so far this was only achieved at the expense of either antifouling or binding capacity. To overcome this limitation, we combined both features into one single monomer: a zwitterionic sulfobetaine, ensuring antifouling capabilities, equipped with a clickable azide moiety which allows for further functionalization. By copolymerizing this monomer together with a standard sulfobetaine, the number of azides (and with that the number of recognition elements) can be tuned depending on the application. First, the clickable azido-monomer was synthesized and characterized, followed by copolymerizing this monomer to yield functionalizable antifouling brushes. The brushes were fully characterized using surface characterization techniques like XPS, contact angle measurements, G-ATR-FTIR and XRR. As a proof of principle, the brushes were subsequently functionalized with biotin via strain-promoted alkyne azide click reactions, which yielded a fully zwitterionic biotin-containing 3D-functionalized coating. The sensing capacity was evaluated by reflectometry using avidin and fibrinogen containing protein solutions. The surfaces showed excellent antifouling properties as illustrated by the complete absence of non-specific fibrinogen binding, while at the same time clear responses were seen for the specific binding of avidin. A great increase in signal-to-noise ratio was observed, even when the amount of functional groups was lowered to 1%, compared to traditional modification of sulfobetaine brushes that rely on a 2D-approach in which only the top-layer can be functionalized. This study was performed on stoichiometric silicon nitride surfaces for future microring resonator based assays, however, this methodology can be transferred to other biosensor platforms which are currently being investigated. The approach presented herein enables a highly efficient strategy for selective binding with retained antifouling properties for improved signal-to-noise ratios in binding assays. The number of recognition units can be adjusted to a specific need, e.g. depending on the size of the analyte to be bound, widening the scope of these functionalizable surface coatings.

Keywords: antifouling, signal-to-noise ratio, surface functionalization, zwitterionic polymer brushes

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1229 Governance Framework for an Emerging Trust Ecosystem with a Blockchain-Based Supply Chain

Authors: Ismael Ávila, José Reynaldo F. Filho, Vasco Varanda Picchi

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The ever-growing consumer awareness of food provenance in Brazil is driving the creation of a trusted ecosystem around the animal protein supply chain. The traceability and accountability requirements of such an ecosystem demand a blockchain layer to strengthen the weak links in that chain. For that, direct involvement of the companies in the blockchain transactions, including as validator nodes of the network, implies formalizing a partnership with the consortium behind the ecosystem. Yet, their compliance standards usually require that a formal governance structure is in place before they agree with any membership terms. In light of such a strategic role of blockchain governance, the paper discusses a framework for tailoring a governance model for a blockchain-based solution aimed at the meat supply chain and evaluates principles and attributes in terms of their relevance to the development of a robust trust ecosystem.

Keywords: blockchain, governance, trust ecosystem, supply chain, traceability

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1228 The Anti-Inflammatory Effects of Nanodiamond Particles and Lipoic Acid on Rats' Cardiovascular System

Authors: Beata Skibska, Andrzej Stanczak, Agnieszka Skibska

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Nanodiamond (ND) is a carbon nanomaterial that has high biocompatibility, and it has a very positive effect on a number of biochemical processes. NDs have great potential in treating multiple inflammation-associated diseases. The purpose of this study was to investigate the anti-inflammatory effect of nanodiamonds and lipoic acid (LA) (as antioxidants) on rats' cardiovascular systems after lipopolysaccharide (LPS) administration. Animal experiments enabled the determination of how nanodiamonds act when applied independently or in combination with lipoic acid. The effect of NDs and LA on C-reactive protein (CRP) levels and heart edema was evaluated. NDs and LA administered after LPS administration attenuated heart edema and significantly decreased the CRP level. The results suggest that NDs and LA play an important role in LPS-induced inflammation in the heart. NDs find new applications in modern biomedical science and biotechnologies.

Keywords: nanodiamonds, lipoic acid, inflammation, cardiovascular system

Procedia PDF Downloads 79
1227 Investigating the Role of Circular RNA GATAD2A on H1N1 Replication

Authors: Tianqi Yu, Yingnan Ding, Yina Zhang, Yulan Liu, Yahui Li, Jing Lei, Jiyong Zhou, Suquan Song, Boli Hu

Abstract:

Circular RNAs (circRNAs) play critical roles in various diseases. However, whether and how circular RNA regulates influenza A virus (IAV) infection is unknown. Here, we studied the role of circular RNA GATA Zinc Finger Domain Containing 2A (circ-GATAD2A) in the replication of IAV H1N1 in A549 cells. Circ-GATAD2A was formed upon H1N1 infection. Knockdown of circ-GATAD2A in A549 cells enhanced autophagy and inhibited H1N1 replication. By contrast, overexpression of circ-GATAD2A impaired autophagy and promoted H1N1 replication. Similarly, knockout of vacuolar protein sorting 34 (VPS34) blocked autophagy and increased H1N1 replication. However, the expression of circ-GATAD2A could not further enhance H1N1 replication in VPS34 knockout cells. Collectively, these data indicated that circ-GATAD2A promotes the replication of H1N1 by inhibiting autophagy.

Keywords: autophagy, circ-GATAD2A, H1N1, replication

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1226 Hepatoprotective Effect of Mycophenolate Mofetil against Tacrolimus Exposure in Rat

Authors: Ferjani Hanen, El Arem Amira, Boussema Ayed Imen, Bacha Hassen

Abstract:

Tacrolimus (TAC), a calcineurin inhibitor, is clinically used as an immunosuppressive agent in the transplant recipient, but its use associated-hepatotoxicity. Mycophenolate mofetil (MMF), an anti-metabolite, is a potent immunosuppressive drug. MMF is not hepatotoxic and is the most common adjunctive immunosuppressant for TAC. The effects of TAC and MMF combination in the liver is still not well understood. This work aimed to investigate their combined effect against in liver in rats Wistar after 24 h. The oral median lethal doses (LD50) of TAC and MMF alone were evaluated in rats are 240 mg/kg and 500 mg/kg respectively. Oral administration of the MMF at 50 mg/kg to male Wistar intoxicated with TAC at 60 mg/kg, demonstrated a significant protective effect by lowering the levels of hepatic markers enzymes (AST, ALT) in the serum rat. MMF attenuated oxidative stress by restoring the activities of SOD, CAT and by reducing the malondialdehyde (MDA) and protein carbonyl levels liver. This study provided evidence that MMF protects rat liver from TAC-induced injury and suggests a most combination use for organ transplantation.

Keywords: tacrolimus, mycophenolate mofetil, combination, liver, rat

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1225 In silico Analysis of Isoniazid Resistance in Mycobacterium tuberculosis

Authors: A. Nusrath Unissa, Sameer Hassan, Luke Elizabeth Hanna

Abstract:

Altered drug binding may be an important factor in isoniazid (INH) resistance, rather than major changes in the enzyme’s activity as a catalase or peroxidase (KatG). The identification of structural or functional defects in the mutant KatGs responsible for INH resistance remains as an area to be explored. In this connection, the differences in the binding affinity between wild-type (WT) and mutants of KatG were investigated, through the generation of three mutants of KatG, Ser315Thr [S315T], Ser315Asn [S315N], Ser315Arg [S315R] and a WT [S315]) with the help of software-MODELLER. The mutants were docked with INH using the software-GOLD. The affinity is lower for WT than mutant, suggesting the tight binding of INH with the mutant protein compared to WT type. These models provide the in silico evidence for the binding interaction of KatG with INH and implicate the basis for rationalization of INH resistance in naturally occurring KatG mutant strains of Mycobacterium tuberculosis.

Keywords: Mycobacterium tuberculosis, KatG, INH resistance, mutants, modelling, docking

Procedia PDF Downloads 314
1224 Evaluation of Pretreatment and Bioactive Compounds Recovery from Chlorella vulgaris

Authors: Marina Stramarkou, Sofia Papadaki, Konstantina Kyriakopoulou, Magdalini Krokida

Abstract:

Nowadays, microalgae represent the diverse branch of microorganism that is used not only in fish farming, but also in food, cosmetics, pharmaceuticals and biofuel production as they can produce a wide range of unique functional ingredients. In the present work, a remarkable microalga Chlorella vulgaris (CV) was selected as a raw material for the recovery of multifunctional extracts. First of all, the drying of raw biomass was examined with freeze-drying showing the best behavior. Ultrasonic-assisted extraction (UAE) using different solvents was applied under the specific optimized conditions. In case of raw biomass, ethanol was the suitable solvent, whereas on dried samples water performed better. The total carotenoid, β-carotene, chlorophyll and protein content in the raw materials, extracts and extraction residues was determined using UV-Vis spectrometry. The microalgae biomass and the extracts were evaluated regarding their antiradical activity using the DPPH method.

Keywords: antioxidant activity, pigments, proteins, ultrasound assisted extraction

Procedia PDF Downloads 330